N-(2,6-dimethylphenyl)-5,6-dihydro-4H-1,3-thiazin-2-amine

Conditions: temp:0.00; pressure:0.00; stir:; See reaction.notes.procedure_details.

Procedure: Rats were anesthetized with a 1.0 ml/kg intramuscular injection of a mixture of ketamine (33 mg/ml), xylazine (1.7 mg/ml), and acepromazine (10 mg/ml) and positioned in a Kopf stereotaxic instrument. A total of 12 μg of 6-OHDA (6 μl volume at a concentration of 2 μg/μl dissolved in 0.9% saline containing 0.2 μg/μl ascorbic acid) were infused at a rate of 1.0 μl/minute and allowed to diffuse for 5 minutes before the infusion cannula was slowly retracted. Infusion coordinates were 4.2 mm posterior to bregma, 1.0 mm lateral from midline, and 7.4 mm ventral to dura.

Conditions: temp:0.00; pressure:0.00; stir:; See reaction.notes.procedure_details.

Procedure: Rats were anesthetized with a 1.0 ml/kg intramuscular injection of a mixture of ketamine (33 mg/ml), xylazine (1.7 mg/ml), and acepromazine (10 mg/ml) and positioned in a Kopf stereotaxic instrument. A total of 12 μg of 6-OHDA (6 μl volume at a concentration of 2 μg/μl dissolved in 0.9% saline containing 0.2 μg/μl ascorbic acid) were infused at a rate of 1.0 μl/minute and allowed to diffuse for 5 minutes before the infusion cannula was slowly retracted. Infusion coordinates were 4.2 mm posterior to bregma, 1.0 mm lateral from midline, and 7.4 mm ventral to dura.

Conditions: temp:0.00; pressure:0.00; stir:; See reaction.notes.procedure_details.

Procedure: Rats (Sprague Dawley) weighing 300–400 gms were purchased from Simonsen laboratories. CVT-3619 was dissolved in DMSO and further diluted in saline. CVT-510 was dissolved in saline. Ketamine was purchased from Fort Dodge Animal Health, Xylazine from Bayer and Acepromazine Maleate from Fermenta Animal Health Co. Metoprolol and propranolol were purchased from SIGMA. Esmolol was obtained from a local pharmacy. further diluted in saline CVT-510 was dissolved in saline

Conditions: temp:0.00; pressure:0.00; stir:; See reaction.notes.procedure_details.

Procedure: Rats (Sprague Dawley) weighing 300–400 gms were purchased from Simonsen laboratories. CVT-3619 was dissolved in DMSO and further diluted in saline. CVT-510 was dissolved in saline. Ketamine was purchased from Fort Dodge Animal Health, Xylazine from Bayer and Acepromazine Maleate from Fermenta Animal Health Co. Metoprolol and propranolol were purchased from SIGMA. Esmolol was obtained from a local pharmacy. further diluted in saline CVT-510 was dissolved in saline

Conditions: temp:0.00; pressure:0.00; stir:; See reaction.notes.procedure_details.

Procedure: 2,6-Dimethylphenylisothiocyanate (15.0 g, 0.093 mole) was added to 3-amino-1-propanol (6.9 g, 0.093 mole) in 60 ml of ether, and the mixture was heated to reflux for 0.5 hr. After the ether was evaporated, 60 ml of concentrated hydrochloric acid was added and refluxing was continued for 0.5 hr. The cooled reaction mixture was treated with 60 ml of water and filtered. The filtrate was made basic with dilute sodium hydroxide, and the precipitate was collected by filtration and washed with water. Recrystallization from ethanol-water (charcoal) gave 16.3 g (80%) of xylazine, m.p. 136°-138° C. (lit. m.p. 140°-141° C.). This step was run three times with yields ranging from 80 to 93%. the mixture was heated to reflux for 0.5 hr After the ether was evaporated 60 ml of concentrated hydrochloric acid was added refluxing The cooled reaction mixture filtered the precipitate was collected by filtration washed with water Recrystallization from ethanol-water (charcoal)

Conditions: temp:0.00; pressure:0.00; stir:; See reaction.notes.procedure_details.

Procedure: Male AR-97Q mice and their normal littermates were castrated or sham-operated via the abdominal route under ketamine-xylazine anesthesia (50 mg/kg ketamine and 10 mg/kg xylazine, i.p.) at 4 weeks of age. Female AR-97Q mice and their littermates were subcutaneously injected 20 μg of testosterone enanthate dissolved in 20 μl of sesame oil weekly from 4 weeks of age until the end of the analysis. The control mice were given the same amount of sesame oil. at 4 weeks

Conditions: temp:0.00; pressure:0.00; stir:; See reaction.notes.procedure_details.

Procedure: 2,6-Dimethylphenylisothiocyanate (15.0 g, 0.093 mole) was added to 3-amino-1-propanol (6.9 g, 0.093 mole) in 60 ml of ether, and the mixture was heated to reflux for 0.5 hr. After the ether was evaporated, 60 ml of concentrated hydrochloric acid was added and refluxing was continued for 0.5 hr. The cooled reaction mixture was treated with 60 ml of water and filtered. The filtrate was made basic with dilute sodium hydroxide, and the precipitate was collected by filtration and washed with water. Recrystallization from ethanol-water (charcoal) gave 16.3 g (80%) of xylazine, m.p. 136°-138° C. (lit. m.p. 140°-141° C.). This step was run three times with yields ranging from 80 to 93%. the mixture was heated to reflux for 0.5 hr After the ether was evaporated 60 ml of concentrated hydrochloric acid was added refluxing The cooled reaction mixture filtered the precipitate was collected by filtration washed with water Recrystallization from ethanol-water (charcoal)

Conditions: temp:0.00; pressure:0.00; stir:; See reaction.notes.procedure_details.

Procedure: Male AR-97Q mice and their normal littermates were castrated or sham-operated via the abdominal route under ketamine-xylazine anesthesia (50 mg/kg ketamine and 10 mg/kg xylazine, i.p.) at 4 weeks of age. Female AR-97Q mice and their littermates were subcutaneously injected 20 μg of testosterone enanthate dissolved in 20 μl of sesame oil weekly from 4 weeks of age until the end of the analysis. The control mice were given the same amount of sesame oil. at 4 weeks

Conditions: temp:50.01; pressure:0.00; stir:; See reaction.notes.procedure_details.

Procedure: 380 ml (373 grams, 4.97 moles) of 3-amino-1-propanol and 450 ml of tetrahydrofuran ("THF") were added to a 5-L flask fitted with a mechanical stirrer, teflon clad thermol-couple and an addition funnel. Then with rapid stirring, 749 grams (4.59 moles) of the 2,6-dimethylphenylisothiocyanate were added dropwise over a 2 hour period. The temperature was kept below 45° C. by controlling the rate of addition and by the use of a cooling bath. The cooling bath was removed, and replaced with a heating mantle. The temperature was raised to 50° C. followed by the addition of 435 ml. concentrated hydrochloric acid. The addition funnel was removed and a distillation head was placed on the flask. Heating was continued with the distillation of THF and water until the internal temperature had reached 95° C., which took about 2 hours. The reaction mixture was then diluted with 2 liters of water and the stirred solution was allowed to cool to room temperature. The aqueous solution was filtered through a 0.2 microfilter. The reaction mixture was made basic by the careful addition of 10% sodium hydroxide. The addition was continued until the pH of the solution was greater than or equal to 12. A solid was collected by filtration, washed with water and partially dryed in the funnel. The solid was then transferred into a flask fitted with a mechanical stirrer and 1 liter of acetone was added. The slurry was stirred for 1 hour using an ice bath to chill the mixture. The product was isolated by filtration, and air dryed to give 750 grams of crude xylazine. The product had a purity of 99% by GC analysis and gave a IR spectrum that was virtually identical to that of a standard sample of xylazine. The 80 MHz 1HNMR spectrum (CDCl3) produced the following results: delta 7.15 (bs, 3, ArH), 4.5 (t, 2, CH2), 3.15 (t, 2, CH2), 2.9-2.0 (m, 8, CH2 and CH3) was in complete agreement with a spectrum of a standard xylazine sample. Furthermore, the mixed melting point of the xylazine sample and a standard showed no depression. fitted with a mechanical stirrer was kept below 45° C. the rate of addition The cooling bath was removed followed by the addition of 435 ml The addition funnel was removed Heating was continued with the distillation of THF and water until the internal temperature had reached 95° C. The reaction mixture was then diluted with 2 liters of water to cool to room temperature The aqueous solution was filtered through a 0.2 microfilter The reaction mixture was made basic by the careful addition of 10% sodium hydroxide The addition A solid was collected by filtration washed with water The solid was then transferred into a flask fitted with a mechanical stirrer 1 liter of acetone was added to chill the mixture The product was isolated by filtration, and air

Conditions: temp:50.01; pressure:0.00; stir:; See reaction.notes.procedure_details.

Procedure: 380 ml (373 grams, 4.97 moles) of 3-amino-1-propanol and 450 ml of tetrahydrofuran ("THF") were added to a 5-L flask fitted with a mechanical stirrer, teflon clad thermol-couple and an addition funnel. Then with rapid stirring, 749 grams (4.59 moles) of the 2,6-dimethylphenylisothiocyanate were added dropwise over a 2 hour period. The temperature was kept below 45° C. by controlling the rate of addition and by the use of a cooling bath. The cooling bath was removed, and replaced with a heating mantle. The temperature was raised to 50° C. followed by the addition of 435 ml. concentrated hydrochloric acid. The addition funnel was removed and a distillation head was placed on the flask. Heating was continued with the distillation of THF and water until the internal temperature had reached 95° C., which took about 2 hours. The reaction mixture was then diluted with 2 liters of water and the stirred solution was allowed to cool to room temperature. The aqueous solution was filtered through a 0.2 microfilter. The reaction mixture was made basic by the careful addition of 10% sodium hydroxide. The addition was continued until the pH of the solution was greater than or equal to 12. A solid was collected by filtration, washed with water and partially dryed in the funnel. The solid was then transferred into a flask fitted with a mechanical stirrer and 1 liter of acetone was added. The slurry was stirred for 1 hour using an ice bath to chill the mixture. The product was isolated by filtration, and air dryed to give 750 grams of crude xylazine. The product had a purity of 99% by GC analysis and gave a IR spectrum that was virtually identical to that of a standard sample of xylazine. The 80 MHz 1HNMR spectrum (CDCl3) produced the following results: delta 7.15 (bs, 3, ArH), 4.5 (t, 2, CH2), 3.15 (t, 2, CH2), 2.9-2.0 (m, 8, CH2 and CH3) was in complete agreement with a spectrum of a standard xylazine sample. Furthermore, the mixed melting point of the xylazine sample and a standard showed no depression. fitted with a mechanical stirrer was kept below 45° C. the rate of addition The cooling bath was removed followed by the addition of 435 ml The addition funnel was removed Heating was continued with the distillation of THF and water until the internal temperature had reached 95° C. The reaction mixture was then diluted with 2 liters of water to cool to room temperature The aqueous solution was filtered through a 0.2 microfilter The reaction mixture was made basic by the careful addition of 10% sodium hydroxide The addition A solid was collected by filtration washed with water The solid was then transferred into a flask fitted with a mechanical stirrer 1 liter of acetone was added to chill the mixture The product was isolated by filtration, and air

Conditions: temp:50.01; pressure:0.00; stir:; See reaction.notes.procedure_details.

Procedure: 380 ml (373 grams, 4.97 moles) of 3-amino-1-propanol and 450 ml of tetrahydrofuran ("THF") were added to a 5-L flask fitted with a mechanical stirrer, teflon clad thermol-couple and an addition funnel. Then with rapid stirring, 749 grams (4.59 moles) of the 2,6-dimethylphenylisothiocyanate were added dropwise over a 2 hour period. The temperature was kept below 45° C. by controlling the rate of addition and by the use of a cooling bath. The cooling bath was removed, and replaced with a heating mantle. The temperature was raised to 50° C. followed by the addition of 435 ml. concentrated hydrochloric acid. The addition funnel was removed and a distillation head was placed on the flask. Heating was continued with the distillation of THF and water until the internal temperature had reached 95° C., which took about 2 hours. The reaction mixture was then diluted with 2 liters of water and the stirred solution was allowed to cool to room temperature. The aqueous solution was filtered through a 0.2 microfilter. The reaction mixture was made basic by the careful addition of 10% sodium hydroxide. The addition was continued until the pH of the solution was greater than or equal to 12. A solid was collected by filtration, washed with water and partially dryed in the funnel. The solid was then transferred into a flask fitted with a mechanical stirrer and 1 liter of acetone was added. The slurry was stirred for 1 hour using an ice bath to chill the mixture. The product was isolated by filtration, and air dryed to give 750 grams of crude xylazine. The product had a purity of 99% by GC analysis and gave a IR spectrum that was virtually identical to that of a standard sample of xylazine. The 80 MHz 1HNMR spectrum (CDCl3) produced the following results: delta 7.15 (bs, 3, ArH), 4.5 (t, 2, CH2), 3.15 (t, 2, CH2), 2.9-2.0 (m, 8, CH2 and CH3) was in complete agreement with a spectrum of a standard xylazine sample. Furthermore, the mixed melting point of the xylazine sample and a standard showed no depression. fitted with a mechanical stirrer was kept below 45° C. the rate of addition The cooling bath was removed followed by the addition of 435 ml The addition funnel was removed Heating was continued with the distillation of THF and water until the internal temperature had reached 95° C. The reaction mixture was then diluted with 2 liters of water to cool to room temperature The aqueous solution was filtered through a 0.2 microfilter The reaction mixture was made basic by the careful addition of 10% sodium hydroxide The addition A solid was collected by filtration washed with water The solid was then transferred into a flask fitted with a mechanical stirrer 1 liter of acetone was added to chill the mixture The product was isolated by filtration, and air

Conditions: temp:0.00; pressure:0.00; stir:; See reaction.notes.procedure_details.

Procedure: The mice were anesthetized with Isoflurane, and then were given an intramuscular injection of 0.1 mL solution of ketamine (50 mg/kg) and xylazine (10 mg/kg). Approximately 10 minutes after the i.m. injection (after the mice could not turn over when placed on their backs), the mice were treated with 12.3 μL of a 10 mg/mL fluasterone/ethanol:limonene:isopropyl myristate solution to give a dose of 5 mg/kg. The mice were anesthetized and on their backs for approximately 30 minutes after application of fluasterone:ethanol:limonene:isopropyl myristate solution. One hour after the application of fluasterone solution, 2 μg of TPA dissolved in 0.2 mL of acetone was applied to the shaved area on the back of each mouse. Approximately 10 minutes